Team:Groningen/9 September 2010

From 2010.igem.org

(Difference between revisions)
(New page: <br> Peter <br> Chaplin detection <br> Samples from the dry freezer were prepared using TFA. <br> The samples were diluted with 1 mL Q-H20 and put on gels: <br> Gel 1: chppt=e deltapt=e bp...)
Line 1: Line 1:
 +
'''Assembly of biobricks in pSB1C3-backbone for submission to parts registry''' ''by Maarten''
 +
 +
O/N cultures of transformants were mini prepped with Sigma Aldrich mini prep kit and control restrictions were performed.
 +
 +
Control restrictions with EcoRI and PstI on constructs:
 +
 +
{| class="wikitable"
 +
|-
 +
| 2 µL construct
 +
|-
 +
| 2 µL Fast Digest Buffer
 +
|-
 +
| 0,5 µL EcoRI Fast Digest
 +
|-
 +
| 0,5 µL PstI Fast Digest
 +
|-
 +
| 15 µL MQ
 +
|}
 +
 +
{| class="wikitable"
 +
|-
 +
! Construct
 +
! Expected band sizes (bp)
 +
|-
 +
| pSB1C3-C
 +
| 2031, 848
 +
|-
 +
| pSB1C3-DC
 +
| 2031, 872
 +
|-
 +
| pSB1C3-E
 +
| 2031, 317
 +
|-
 +
| pSB1C3-H
 +
| 2031, 302
 +
|-
 +
| pSB1C3-Sortase
 +
| 2031, 689
 +
|-
 +
| pSB1C3-RFP
 +
| 2031, 1069
 +
|}
 +
<br>
<br>
Peter
Peter

Revision as of 21:22, 7 October 2010

Assembly of biobricks in pSB1C3-backbone for submission to parts registry by Maarten

O/N cultures of transformants were mini prepped with Sigma Aldrich mini prep kit and control restrictions were performed.

Control restrictions with EcoRI and PstI on constructs:

2 µL construct
2 µL Fast Digest Buffer
0,5 µL EcoRI Fast Digest
0,5 µL PstI Fast Digest
15 µL MQ
Construct Expected band sizes (bp)
pSB1C3-C 2031, 848
pSB1C3-DC 2031, 872
pSB1C3-E 2031, 317
pSB1C3-H 2031, 302
pSB1C3-Sortase 2031, 689
pSB1C3-RFP 2031, 1069


Peter
Chaplin detection
Samples from the dry freezer were prepared using TFA.
The samples were diluted with 1 mL Q-H20 and put on gels:
Gel 1: chppt=e deltapt=e bpte e1pte enpte chste deltaste bste e1ste
Gel 2: enste deltapt=4 bpt=4 e1pt=4 enpt=4 deltast=4 bst=4 ent=4
Gel 3: est=4 deltadt=e bdt=e e1dt=e endt=e 1 2 3
Gel 4: 4 e1dt=e bdt=e e1nt=e e1pt=e bpt=e e1st=e b1st=e